The HADHB Knockout A-549 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population that serves as a loss-of-function model for the HADHB gene in the human A-549 lung adenocarcinoma cell line. This polyclonal product is generated via CRISPR/Cas9-mediated gene disruption, yielding a heterogeneous mixture of edited cells with targeted inactivation of HADHB. The pooled format captures diverse editing events, reducing clonal bias and allowing robust assessment of gene function.
The A-549 cell line, originally derived from a lung carcinoma of a 58-year-old Caucasian male, exhibits adherent epithelial morphology and is a widely used model for lung adenocarcinoma. It harbors a KRAS mutation and retains alveolar type II epithelial features, making it suitable for investigating cancer metabolism, drug resistance, and signaling pathways.
HADHB encodes the ??-subunit of the mitochondrial trifunctional protein (MTP), which performs the long-chain 3-ketoacyl-CoA thiolase step in the fatty acid ??-oxidation spiral. Working in complex with HADHA, it processes substrates supplied by acyl-CoA synthetase, CPT1, CPT2, and the carnitine shuttle. The catalytic activity is transcriptionally regulated by PPARA and PPARD, and stimulated by hormonal signals including glucagon and epinephrine. The reaction generates acetyl-CoA, NADH, FADH2, and short-chain acyl-CoA. CRISPR/Cas9-mediated disruption of HADHB abolishes long-chain fatty acid catabolism, causing accumulation of long-chain acyl-CoA intermediates and mitochondrial energy deficits.
In A-549 lung adenocarcinoma cells, HADHB knockout allows dissection of the role of fatty acid ??-oxidation in tumor metabolism. Cancer cells often exhibit altered lipid utilization for growth and survival; loss of HADHB may force a metabolic shift toward glycolysis and glutamine dependence, potentially sensitizing cells to metabolic stress or therapies targeting these pathways. This model also provides insights into inherited mitochondrial disorders such as trifunctional protein deficiency, which is associated with cardiomyopathy, hypoglycemia, and rhabdomyolysis, and can be used to screen for modulators of fatty acid oxidation defects.
Typical research applications include metabolic disease modeling, cancer metabolism studies, fatty acid oxidation research, and drug screening for metabolic disorders. Representative assays include western blotting and RT-qPCR for expression validation, fatty acid oxidation assays using radiolabeled palmitate, metabolomics profiling, oxygen consumption rate measurement, lactate production assays, and immunofluorescence. This polyclonal knockout cell population is also amenable to high-throughput screening. For further information, please contact Ascent Research.
